PROJECT SUMMARY More than 80,000 laboratories globally are working on biological sample processing, with a rapidly growing focus on next-generation sequencing (NGS) technologies. The milestone of the $1000 human genome sequence has established NGS as the gold standard technology for personalized/precision medicine and diagnostics. Genetic testing kids are now as readily available as pregnancy test kids, and advances in genetic analysis, gene therapy, and genetic modification promise in the 21st century to parallel the development of transistors and integrated circuits in the 20th century. The challenge is that technology for preparing biological samples for sequencing has not maintained a similar rapid advancement to gene sequencing. Fragmentation of DNA randomly without bias is an essential, but difficult step in the preparation of genomic DNA for NGS applications. A narrow range of DNA fragment lengths is required for sequencing, and inefficient or inconsistent fragmentation will result in incomplete or excluded reads, corrupting the sequencing. Acoustic sonication, which fragments DNA through cavitation, is the gold standard for DNA fragmentation. However, our customer discover interviews have reinforced that this is a significant pain point in their sample preparation process due to its low throughput and challenging consistency, particularly for industry developing large-scale sequencing efforts. Our NCI IMAT-funded academic co-founders have invented a novel approach that overcomes throughput and consistency bottlenecks in sample preparation for next-generation sequencing (NGS) technologies, based on our preliminary data. We have invented a novel nanodroplet reagent that enhances cavitation from an acoustic sonicator, and greatly improves consistency and reduces time and acoustic power requirements for DNA fragmentation. Triangle Biotechnology has licensed the patents describing the cavitation enhancing technology. With this SBIR, we will establish feasibility of this novel reagent, and perform key research, development, and proof of concept studies required prior to commercialization. Success of this project will enable a novel commercial solution for addressing market pain points: substantially improving throughput and consistency of DNA fragmentation for NGS sample preparation while reducing cost.